Disease detection and treatment based on phenylacetyl glutamine levels

Abstract

The present invention relates to systems, kits, and methods for identifying subjects with increased levels of phenylacetyl glutamine (PAG) or the combination of PAG and trimethylamine-n-oxide (TMAO) and/or N6-trimethyl-lysine (TML), and/or PSA, pp and/or betaine, and/or choline, as well as methods of determining risk of disease (e.g., CVD, heart failure, asthma, diabetes, thrombosis, and lethal prostate cancer) based on such levels. In certain embodiments, the subjects are free of chronic kidney disease and/or have type II diabetes. In particular embodiments, subjects are treated with a therapeutic, such as a beta-adrenergic blocking agent, an alpha 2 adrenergic receptor agonist, an alpha 2 adrenergic receptor antagonist, an antibiotic or antibiotic cocktail, or a prostate cancer therapeutic. In certain embodiments, the subject is treated with a procedure such as brachytherapy, radiation therapy, or prostatectomy.

What is claimed is: 
     
         1 . A method performing an activity based on the level of at least phenylacetyl glutamine (PAG) in a sample from a subject comprising:
 a) determining the level of at least one compound in a sample from a subject, wherein said at least one compound comprises PAG, and wherein optionally said subject is chronic kidney disease (CKD) free and/or has type II diabetes; and   b) performing at least one of the following activities:
 i) identifying increased levels of said at least one compound in said sample compared to control levels, and treating said subject with:
 A) a first agent or first procedure that treats cardiovascular disease (CVD), asthma, heart failure, and/or thrombosis, wherein said subject is CKD free and/or has diabetes; 
 B) a second agent selected from: a beta-adrenergic blocking agent, an alpha 2 adrenergic receptor agonist, an alpha 2 adrenergic receptor antagonist, and an antibiotic or antibiotic cocktail; and/or 
 C) a third agent or second procedure selected from: a prostate cancer therapeutic, brachytherapy, radiation therapy, and prostatectomy, 
 
 ii) generating and/or transmitting a report that indicates that levels of said at least one compound are elevated in said sample compared to control levels, and that said subject has, or is at risk for, CVD, thrombosis, asthma, lethal prostate cancer, and/or heart failure and is treatable by: A) said beta-adrenergic blocking agent, and/or B) said alpha 2 adrenergic receptor agonist, and/or C) said an alpha 2 adrenergic receptor antagonist and/or D) said antibiotic or antibiotic cocktail; and/or E) said third agent or second procedure; 
 iii) generating and/or transmitting a report that indicates that levels of said at least one compound are elevated in said sample compared to control levels, and that said subject is in need of said first agent, said second agent, and/or said first procedure, and/or said third agent or second procedure; 
 iv) generating and/or transmitting a report for a subject that is CKD free and/or has diabetes, wherein said report indicates that levels of said at least one compound are elevated in said sample compared to control levels, and that said subject has, or is at risk for, cardiovascular disease, thrombosis, lethal prostate cancer, and/or heart failure; and 
 v) characterizing said subject who is CKD free and/or has diabetes as having, or at risk for, CVD, thrombosis, and/or heart failure, or characterizing said subject as having, or at risk for, lethal prostate cancer, based on finding elevated levels of said at least one compound in said sample compared to a control levels. 
   
     
     
         2 . The method of  claim 1 , wherein said at least one compound further comprises trimethylamine-n-oxide (TMAO). 
     
     
         3 . The method of  claim 1 , wherein: i) said subject has heart failure, asthma, or cardiovascular disease, and wherein said at least one compound further comprises N6-trimethyl-lysine (TML) and/or TMAO, or ii) said subject has prostate cancer, and said at least one compound further comprises PSA, choline, and/or betaine. 
     
     
         4 . The method of  claim 1 , wherein said at least one compound further comprises TMAO and TML. 
     
     
         5 . The method of  claim 1 , wherein said sample is selected from the group consisting of:
 a plasma sample, a serum sample, a whole blood sample, and a urine sample.   
     
     
         6 . The method of  claim 1 , wherein said determining comprises detecting said at least one compound with an analytical device selected from: a mass spectrometer, NMR spectrometer, and a UV/Vis spectrometer. 
     
     
         7 . The method of  claim 1 , wherein said determining comprises contacting said sample with an anti-PAG antibody or PAG binding fragment thereof. 
     
     
         8 . The method of  claim 1 , wherein said a beta-adrenergic blocking agent is selected from: acebutolol hydrochloride, atenolol, betaxolol hydrochloride, bisoprolol fumarate, carteolol hydrochloride, esmolol hydrochloride, metoprolol, penbutolol sulfate, nadolol, nebivolol, pindolol, propranolol, timolol maleate, sotalol hydrochloride, carvedilol, and labetalol hydrochloride. 
     
     
         9 . The method of  claim 1 , wherein said alpha 2 adrenergic receptor agonist is selected from: Clonidine, Dexmedetomidine, Fadolmidine, Guanfacine, Guanabenz, Guanoxabenz, Guanethidine, Xylazine, Tizanidine, Medetomidine, Methyldopa, Methylnorepinephrine, Norepinephrine, (R)-3-nitrobiphenyline, amitraz, Detomidine, Lofexidine,and Medetomidine. 
     
     
         10 . The method of  claim 1 , wherein said alpha 2 adrenergic receptor antagonist is selected from: atipamezole, efaroxan, idazoxan, yohimbine, rauwolscine, and phentolamine. 
     
     
         11 . The method of  claim 1 , wherein said first agent is selected from the group consisting of: an anticoagulant, an antiplatelet agent, an ACE Inhibitor, an angiotensin II receptor blocker, an angiotensin-receptor neprilysin inhibitor, a calcium channel blocker, a cholesterol-lowering medication, a statin, a digitalis preparation, a diuretic, and a vasodilator. 
     
     
         12 . The method of  claim 1 , wherein said antibiotic is at least one antibiotic selected from the group consisting of: metronidazole, ciprofloxacin, neomycin, vancomycin, amoxicillin, and a broad spectrum antibiotic; and wherein said subject does not have an active infection. 
     
     
         13 . The method of  claim 1 , wherein said subject is a human with, or at risk for, prostate cancer, cardiovascular disease, asthma, or heart failure. 
     
     
         14 . A method of treatment comprising:
 a) identifying a subject as having increased levels of at least one compound compared to control levels, wherein said at least one compound comprises phenylacetyl glutamine (PAG); and   b) treating said subject with at least one of the following:
 i) a first agent or procedure that treats cardiovascular disease (CVD), heart failure, asthma, and/or thrombosis, wherein said subject is chronic kidney disease (CKD) free and/or has diabetes, 
 ii) a second agent selected from: a beta-adrenergic blocking agent, an alpha 2 adrenergic receptor agonist, and an alpha 2 adrenergic receptor antagonist, 
 iii) an antibiotic or antibiotic cocktail, wherein said subject does not have an active infection; 
 iv) a third agent or second procedure selected from: a prostate cancer therapeutic, brachytherapy, radiation therapy, and prostatectomy. 
   
     
     
         15 . The method of  claim 14 , wherein said identifying comprises receiving a report that said subject has increased PAG levels compared to a control. 
     
     
         16 . The method of  claim 14 , wherein said at least one compound further comprises PSA, choline, and/or betaine, and said subject has prostate cancer. 
     
     
         17 . The method of  claim 14 , wherein said subject is a human having heart failure, asthma, cardiovascular disease, or prostate cancer. 
     
     
         18 . The method of  claim 17 , wherein said a beta-adrenergic blocking agent is selected from: acebutolol hydrochloride, atenolol, betaxolol hydrochloride, bisoprolol fumarate, carteolol hydrochloride, esmolol hydrochloride, metoprolol, penbutolol sulfate, nadolol, nebivolol, pindolol, propranolol, timolol maleate, sotalol hydrochloride, carvedilol, and labetalol hydrochloride. 
     
     
         19 . The method of  claim 14 , wherein said alpha 2 adrenergic receptor agonist is selected from: Clonidine, Dexmedetomidine, Fadolmidine, Guanfacine, Guanabenz, Guanoxabenz, Guanethidine, Xylazine, Tizanidine, Medetomidine, Methyldopa, Methylnorepinephrine, Norepinephrine, (R)-3-nitrobiphenyline, amitraz, Detomidine, Lofexidine,and Medetomidine. 
     
     
         20 . The method of  claim 14 , wherein said alpha 2 adrenergic receptor antagonist is selected from: atipamezole, efaroxan, idazoxan, yohimbine, rauwolscine, and phentolamine. 
     
     
         21 . The method of  claim 14 , wherein said first agent is selected from the group consisting of: an anticoagulant, an antiplatelet agent, an ACE Inhibitor, an angiotensin II receptor blocker, an angiotensin-receptor neprilysin inhibitor, a calcium channel blocker, a cholesterol-lowering medication, a statin, a digitalis preparation, a diuretic, and a vasodilator. 
     
     
         22 . The method of  claim 14 , wherein said antibiotic is at least one antibiotic selected from the group consisting of: metronidazole, ciprofloxacin, neomycin, vancomycin, amoxicillin, and a broad spectrum antibiotic; and wherein said subject does not have an active infection. 
     
     
         23 . The method of  claim 14 , wherein said subject has heart failure, asthma, or cardiovascular disease, and wherein said at least one compound further comprises trimethylamine-n-oxide (TMAO) and/or N6-trimethyl-lysine (TML) 
     
     
         24 . A method of detecting at least one compound in a sample comprising:
 a) obtaining a sample, wherein said sample is from a human subject who is optionally chronic kidney disease (CKD) free and/or has diabetes; and   b) treating said sample under conditions such that the concentration of at least one compound is determined, wherein said at least one compound comprises phenylacetyl glutamine (PAG).   
     
     
         25 . The method of  claim 24 , wherein said concentration of said PAG in said sample is determined by mass spectrometry. 
     
     
         26 . The method of  claim 24 , wherein said wherein said subject has heart failure, asthma, cardiovascular disease, or prostate cancer. 
     
     
         27 . The method of  claim 24 , wherein said at least one compound further comprises trimethylamine-n-oxide (TMAO) and/or N6-trimethyl-lysine (TML) and/or PSA and/or choline, and/or betaine. 
     
     
         28 . A system or kit comprising:
 a) a report for a subject with cardiovascular disease, heart failure, asthma, thrombosis, and/or prostate cancer, wherein said report indicates that said patient has elevated levels of at least one compound, wherein said at least one compound comprises phenylacetyl glutamine (PAG); and   b) at least one of the following:
 i) a first agent that treats CVD, heart failure, and/or thrombosis, wherein said subject is chronic kidney disease (CKD) free and/or has diabetes, 
 ii) a second agent selected from: a beta-adrenergic blocking agent, an alpha 2 adrenergic receptor agonist, and an alpha 2 adrenergic receptor antagonist, 
 iii) an antibiotic or antibiotic cocktail, wherein said subject does not have an active infection, and 
 iv) a prostate cancer therapeutic. 
   
     
     
         29 . The system of  claim 28 , wherein said subject is a human. 
     
     
         30 . The system of  claim 28 , wherein said a beta-adrenergic blocking agent is selected from: acebutolol hydrochloride, atenolol, betaxolol hydrochloride, bisoprolol fumarate, carteolol hydrochloride, esmolol hydrochloride, metoprolol, penbutolol sulfate, nadolol, nebivolol, pindolol, propranolol, timolol maleate, sotalol hydrochloride, carvedilol, and labetalol hydrochloride. 
     
     
         31 . The system of  claim 28 , wherein said alpha 2 adrenergic receptor agonist is selected from: Clonidine, Dexmedetomidine, Fadolmidine, Guanfacine, Guanabenz, Guanoxabenz, Guanethidine, Xylazine, Tizanidine, Medetomidine, Methyldopa, Methylnorepinephrine, Norepinephrine, (R)-3-nitrobiphenyline, amitraz, Detomidine, Lofexidine,and Medetomidine. 
     
     
         32 . The system of  claim 28 , wherein said alpha 2 adrenergic receptor antagonist is selected from: atipamezole, efaroxan, idazoxan, yohimbine, rauwolscine, and phentolamine. 
     
     
         33 . The system of  claim 28 , wherein said first agent is selected from the group consisting of: an anticoagulant, an antiplatelet agent, an ACE Inhibitor, an angiotensin II receptor blocker, an angiotensin-receptor neprilysin inhibitor, a calcium channel blocker, a cholesterol-lowering medication, a statin, a digitalis preparation, a diuretic, and a vasodilator. 
     
     
         34 . The system of  claim 28 , wherein said antibiotic is at least one antibiotic selected from the group consisting of: metronidazole, ciprofloxacin, neomycin, vancomycin, amoxicillin, and a broad spectrum antibiotic; and wherein said subject does not have an active infection. 
     
     
         35 . The system of  claim 28 , wherein said at least one compound further comprises trimethylamine-n-oxide (TMAO) and/or N6-trimethyl-lysine (TML) and/or PSA and/or choline, and/or betaine. 
     
     
         36 . A method of detecting phenylacetyl glutamine (PAG) in a serum or plasma sample comprising:
 a) obtaining a plasma or serum sample, wherein said plasma or serum sample is from a human subject who is chronic kidney disease (CKD) free and/or has type II diabetes;   b) introducing at least a portion of said plasma or serum sample into an analytical device under conditions such that the concentration of PAG present in said plasma or serum sample is determined,   wherein said analytical device comprises: i) an NMR spectrometer, a UVNis spectrometer, or a mass spectrometer, ii) equipment to provide physical separation of said PAG prior to determining said concentration, and iii) an anti-PAG antibody based detection system; and   c) graphically displaying said subject's risk of having a disease as higher than normal if said level of PAG in said plasma or serum sample is higher than a first minimum value, wherein said minimum value is at least 4.9 μM or at least 3.8 μM, wherein said disease is selected from the group consisting of heart failure, asthma, cardiovascular disease, or thrombosis.   
     
     
         37 . The method of  claim 36 , further comprising determining the level of trimethylamine N-oxide (TMAO) in a sample from a subject, and graphically displaying said subject's risk of said disease as higher than normal when said TMAO level is higher than a second minimum value, wherein said second minimum value is at least 2.2 μM. 
     
     
         38 . The method of  claim 36 , wherein said second minimum value is at least 2.5 μM. 
     
     
         39 . The method of  claim 36 , wherein said second minimum value is at least 3.0 μM. 
     
     
         40 . The method of  claim 36 , wherein said second minimum value is at least 4.0 μM. 
     
     
         41 . The method of  claim 36 , wherein said second minimum value is at least 5.0 μM. 
     
     
         42 . The method of  claim 36 , further comprising determining the level of N6-trimethyl-lysine (TML) in a sample from a subject, and graphically displaying said subject's risk of said disease as higher than normal when said TML level is higher than a second minimum value, wherein said second minimum value is at least 0.4 μM. 
     
     
         43 . The method of  claim 42 , wherein said second minimum value is at least 0.5 μM. 
     
     
         44 . The method of  claim 42 , wherein said second minimum value is at least 0.6 μM. 
     
     
         45 . The method of  claim 42 , wherein said second minimum value is at least 0.7 μM. 
     
     
         46 . The method of  claim 42 , wherein said second minimum value is at least 0.8 μM. 
     
     
         48 . A method of detecting at least three compounds in a sample comprising:
 a) obtaining a sample, wherein said sample is from a human subject; and   b) treating said sample under conditions such that the concentration of at least the following three compounds is determined: phenylacetyl glutamine (PAG), N6-trimethyl-lysine (TML), and trimethylamine N-oxide (TMAO).   
     
     
         49 . The method of  claim 48 , further comprising: c) graphically displaying said subject's risk of having a disease as higher than normal if all three are present:
 i) said level of PAG in said sample is higher than a control PAG value from the general population or disease free group;   ii) said TMAO level in said sample is higher than a control TMAO value from the general population or a disease free group; and   iii) said TML level is in said sample is higher than a control TML value from the general population or a disease free group; and   wherein said disease is selected from the group consisting of heart failure, cardiovascular disease, kidney disease, asthma, or thrombosis.   
     
     
         50 . The method of  claim 48 , further comprising: c) graphically displaying said subject's risk of having a disease as higher than normal if all three are present:
 i) said level of PAG in said sample is higher than a first minimum value, wherein said minimum value is at least 3.9 or 4.9 M,   ii) said TMAO level is higher than a second minimum value, wherein said second minimum value is at least 2.2 or 5.0 μM; and   iii) said TML level is higher than a third minimum value, wherein said second minimum value is at least 0.4 μM or 0.6; and   wherein said disease is selected from the group consisting of heart failure, cardiovascular disease, kidney disease, asthma, or thrombosis.   
     
     
         51 . A method of detecting phenylacetyl glutamine (PAG) in a serum or plasma sample comprising:
 a) obtaining a plasma or serum sample, wherein said plasma or serum sample is from a human subject who is optionally chronic kidney disease (CKD) free and/or has type II diabetes;   b) introducing at least a portion of said plasma or serum sample into an analytical device under conditions such that the concentration of PAG present in said plasma or serum sample is determined,   wherein said analytical device comprises: i) an NMR spectrometer, a UVNis spectrometer, or a mass spectrometer, ii) equipment to provide physical separation of said PAG prior to determining said concentration; and iii) an anti-PAG antibody based detection system; and   c) graphically displaying said subject's risk of having a disease as higher than normal if said level of PAG in said plasma or serum sample is higher than a first minimum value, wherein said minimum value is at least 1.0 μM or at least 3.0 μM, wherein said disease is selected from the group consisting of prostate cancer and lethal prostate cancer.   
     
     
         52 . The method of  claim 51 , further comprising determining the level of choline in a sample from a subject, and graphically displaying said subject's risk of said disease as higher than normal when said choline level is higher than a second minimum value, wherein said second minimum value is at least 11.0 μM. 
     
     
         53 . The method of  claim 51 , wherein said second minimum value is at least 13.0 μM. 
     
     
         54 . The method of  claim 51 , wherein said second minimum value is at least 14.0 μM. 
     
     
         55 . The method of  claim 51 , wherein said second minimum value is at least 15.0 μM. 
     
     
         56 . The method of  claim 51 , wherein said second minimum value is at least 17.0 μM. 
     
     
         57 . The method of  claim 51 , further comprising determining the level of betaine in a sample from a subject, and graphically displaying said subject's risk of said disease as higher than normal when said betaine level is higher than a second minimum value, wherein said second minimum value is at least 35.6 μM. 
     
     
         58 . The method of  claim 57 , wherein said second minimum value is at least 42.8 μM. 
     
     
         59 . The method of  claim 57 , wherein said second minimum value is at least 54.0 μM. 
     
     
         60 . The method of  claim 57 , wherein said second minimum value is at least 54.7 μM. 
     
     
         61 . The method of  claim 57 , wherein said second minimum value is at least 55.0 μM. 
     
     
         62 . A method of detecting at least three or four compounds in a sample comprising:
 a) obtaining a sample, wherein said sample is from a human subject; and   b) treating said sample under conditions such that the concentration of at least the following three compounds is determined: phenylacetyl glutamine (PAG), choline, and betaine, and optionally a fourth compound: prostate specific antigen (PSA).   
     
     
         63 . The method of  claim 62 , further comprising: c) graphically displaying said subject's risk of having a disease as higher than normal if all three are present:
 i) said level of PAG in said sample is higher than a control PAG value from the general population or disease free group;   ii) said choline level in said sample is higher than a control choline value from the general population or a disease free group; and   iii) said betaine level is in said sample is higher than a control TML value from the general population or a disease free group; and   iv) and optionally, said PSA level in said sample is higher than a control PSA value from the general population or disease free group; and   wherein said disease is selected from the group consisting of prostate cancer and lethal prostate cancer.   
     
     
         64 . The method of  claim 63 , further comprising: c) graphically displaying said subject's risk of having a disease as higher than normal if all three are present:
 i) said level of PAG in said sample is higher than a first minimum value, wherein said minimum value is at least 3.9 or 4.9 M,   ii) said choline level is higher than a second minimum value, wherein said second minimum value is at least 11.0 or 13.0 μM; and   iii) said betaine level is higher than a third minimum value, wherein said second minimum value is at least 35.6 μM or 42.8; and   wherein said disease is selected from the group consisting of prostate cancer and lethal prostate cancer.